Vol. 303, Issue 1, 74-81, October 2002

Potent Suppression of Proliferation of A10 Vascular Smooth Muscle Cells by Combined Treatment with Lovastatin and 3-Allylfarnesol, an Inhibitor of Protein Farnesyltransferase

Raymond R. Mattingly , Richard A. Gibbs, Raymond E. Menard and John J. Reiners, Jr.

Department of Pharmacology (R.R.M, R.E.M.), Wayne State University, Detroit, Michigan; Programs in Molecular Biology and Genetics (R.R.M.) and Proteases (J.J.R.), Barbara Ann Karmanos Cancer Institute, Detroit, Michigan; Department of Medicinal Chemistry and Molecular Pharmacology (R.A.G.), College of Pharmacy and Pharmacal Sciences, Purdue University, West Lafayette, Indiana; and Institute of Environmental Health Sciences (J.J.R.), Wayne State University, Detroit, Michigan

Statins, which inhibit 3-hydroxy-3-methylglutaryl-CoA reductase and thus the synthesis of cholesterol, are remarkably effective in the treatment of cardiovascular disease. In addition to their favorable effect on lipid profile, these drugs may also prevent the proliferation of vascular smooth muscle that is characteristic of atherosclerosis. We hypothesize that statins prevent the post-translational prenylation, and thus inhibit the function, of critical small GTPases in vascular smooth muscle cells. We have therefore assayed the effect of lovastatin on both the growth of A10 vascular smooth muscle cells and the status of their Ras and RhoB proteins. We find that 1 µM lovastatin potently inhibits the proliferation of A10 cultures, and higher concentrations (3 µM) induce apoptosis. We have also tested the effect of 3-allylfarnesol (3-alFOH), an inhibitor of farnesyl transferase (FTI). The data show that although 10 µM 3-alFOH is required for a cytostatic effect, the action of 3 µM 3-alFOH can be greatly potentiated by even nanomolar levels of lovastatin. We also find that lovastatin and 3-alFOH exhibit synergism to cause the up-regulation and relocalization of RhoB from the membrane to cytosolic compartments. This relocalization of RhoB, which is presumed to reflect an inhibition of its prenylation, correlates with the proapoptotic activities of combined 3-alFOH and lovastatin treatment. These data suggest that RhoB may be a valuable pharmacological target in cardiovascular disease, and that combinations of statins and certain FTIs may be of value in treatment of disorders that are characterized by excess cell proliferation.